JPS6327432A - Local methotrexate preparation for treating hyperproliferative epidermal disease and remeady - Google Patents

Abstract

Topical compositions containing methotrexate or its analogs and an effective skin penetration agent, as well as methods of its use in the topical treatment of hyperproliferative epithelial diseases, such as psoriasis.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to topical preparations containing methotrexate or analogues thereof in the treatment of hyperproliferative epithelial diseases, such as insensitivity.

BACKGROUND OF THE INVENTION The invention described herein was developed by the National Institutes of Health (the Nat.
ional In5 postures of Heart
h), U.S. Department of Health and Human Services (U.S., Depar.
tment of Health and Human
Research direction was carried out under the permission or approval of

Epithelial diseases (epidermal and mucosal) are a serious health problem. Some of the specific issues are.

It is characterized by an abnormal (increased) rate of cell replacement and is described here as a hyperproliferative epithelial disease. Examples of hyperproliferative epithelial diseases include dry skin.

Skin tumors of cutaneous origin (basic cell carcinoma, squamous cell carcinoma.

melanoma, mycosis, Bowen's disease), viral diseases (warts, herpes simplex, condyloma acuminata); 9 premalignant and malignant diseases of the female reproductive tract (cervix, vagina, vulva); There are premalignant and malignant diseases of mucosal cells (oral cavity, bladder, rectum).

The most common of these diseases is dry skin. Recent estimates suggest that in the United States, 100 to 300
It has been shown that 10,000 people suffer from dry drunkenness, and approximately 150,000 to 250,000 new cases are reported every year. The prevalence of Qianfu in the United States is similar to the prevalence in other countries, with a population of 2.
% and 4%. Therefore, there is a great need for effective treatments.

Recently, a variety of treatments have been used to treat dryness. This treatment includes dialysis, chemotherapy (local and systemic)
and photochemotherapy (local and systemic). Local chemotherapy is perhaps the most widely used. Use reagents.

Currently, the most severe cases of Qiantang are treated with systemic photochemotherapy or PUVA (psoralen + OVA). However, with such treatments there is a risk of directly altering the DNA structure of the patient's cells. Therefore, such treatment methods are potentially mutagenic and carcinogenic (R,S,5ter
(1979), New England Medical Journal, 3007809-813), systemic chemotherapy using methotrexate (MTX) is also used effectively in the treatment of severe or extensive xerosis. . MTX is administered orally or by injection. However, if used chronically,
Causes significant side effects (especially liver damage) (Wein
Stein, (1977) + Ann Tnt Me
d: 199).

The majority of patients with phlegm have minimal or moderate involvement. Therefore, the risks associated with systemic chemotherapy with pUvA or MTX are not warranted. Such patients are generally treated locally with steroids. What is the form of treatment?

Not effective either at the beginning or at the end. Therefore.

For the majority of patients suffering from proliferative skin diseases.

Effective and safe local treatments are needed.

MTX has been demonstrated to be an effective systemic chemotherapeutic agent in the treatment of drywall (VanScott
et al+ (1964) Arch Derm
89:550; Reese et al. (1967
)^rch Derm 95:2), if it is MTX.

It is suggested that it can be developed into an effective chemotherapeutic agent for local delivery. Topical application of MTX is highly desirable. This is because the harmful side effects of systemic administration are reduced because the drug is applied only to the painful tissue, and non-painful tissues, such as the liver, are exposed to little or no drug.

However, some prior art studies applying MTX topically have reported that this drug is not effective (See Weinstein et al.
al.

(1981) 8arch Derm 11:388; Weinstein inPharm
acolo and the 5kin; Adva
ces in Biol.

of the 5kin, Vol, XHpp,
287-202 (W, Montana.

RB Stoughton & EJ van 5co.
tt eds, 1969); van Scott e.
tal, (1959) J Invest De
rm 33:357; Nurse, (1963)
Arch Derm 87:168;
etal, (1969) Arch Derm
100: 99; Stewart et al. (1
972) Arch Derm 106:357). For example, in Einstein et al. (1981), methotrexate was clinically enhanced in a vehicle containing a permeation-enhancing agent (C-10 methyl sulfoxide) at 2.5%. 6 patients with 0.1% and 0.5%
Treatment was administered daily for 9 days with a vehicle formulation containing MTX. No significant improvement was seen in any patient.

Other studies have shown that two topical MTX preparations have slight percutaneous penetration through human skin in vitro and in vivo (MsCulloughet a
l+ (1976) J Invest Derm
66:103; Stewart et al+ (19
72) u anal L: Comaish et aL (1
969)...Anal, ) 6 Therefore, MTX is (
a) Does not penetrate the skin well; Q)) Acts systemically at sites remote from the affected tissue. or (C) convert to an active form at a site distant from the afflicted tissue. (Weinstein
et al (1976) J Invest Der.
See M 67:26).

Further interesting studies on the mechanism of action of MTX include + (Weinstein et al. (1971)
) Arch Derm104: 236 and N
ewberger eL al, (1978) J I
nvestDerm 70:183).

SUMMARY OF THE INVENTION It is an object of the present invention to provide an effective therapy for the treatment of hyperproliferative epithelial diseases.

It is also an object of the invention to provide a topical preparation for the treatment of hyperproliferative epithelial diseases.

Another object of the invention is to provide a therapy for the treatment of hyperproliferative epithelial diseases that avoids the disadvantages associated with current PUV therapy or systemic chemotherapy.

Yet another object of the invention is to provide a localized treatment for hyperproliferative epithelial diseases.

Yet another object of the present invention is to provide a topical formulation that can enhance the transdermal permeability of methotrexate or its derivatives.

These and other objects and advantages of the present invention will be readily apparent to those skilled in the art from the embodiments described below.

In one embodiment, the present invention provides compositions for topical application to treat hyperproliferative epithelial diseases.

The composition contains: (a) an effective amount of a skin permeation reagent that permeates methotrexate into the basal epidermal cells of a mammal to inhibit DNA synthesis in the basal epidermal cells; a topical carrier comprising in sufficient amount, and (b) a therapeutically effective amount of an agent selected from the group consisting of methotrexate and methotrexate analogs. here,
The skin permeation reagent has the following formula: where R' is selected from the group consisting of H1 and alkyl having 1 to 4 carbon atoms.

R is selected from the group consisting of alkyl of 1 to 18 carbon atoms, and phenyl, and n is 0 or 1-1
It is a positive integer of 0.

In another embodiment, the invention provides a method of treatment for the treatment of hyperproliferative epithelial diseases, comprising:
A therapeutically effective amount of methotrexate or a methotrexate analog is applied to an area of the mammal's skin that is affected by the hyperproliferative epithelial disease.

It is administered locally. wherein the administration results in sufficient methotrexate or methotrexate analog to penetrate the basal epidermal cells of the mammal in an amount sufficient to inhibit DNA synthesis in the basal epidermal cells.

The method of treating a hyperproliferative epithelial disease of the present invention is a method for therapeutically treating a hyperproliferative epithelial disease in a mammal, comprising: providing the above composition; Applications to mammals with proliferative epithelial disease pain.

Other implementations of the invention! IE, will be readily apparent to those skilled in the art from the following disclosure.

DETAILED DESCRIPTION OF THE INVENTION Methotrexate and its analogs have long been known to have therapeutic activity against hyperproliferative epithelial diseases such as xerosis. Although MTX is effective when administered systemically, exposing the entire body to the drug has one serious side effect. Therefore, the prior art tested methotrexate in two topical formulations. However, these trials did not produce a clear therapeutic effect. This raised the possibility that methotrexate's activity is linked to its mode of administration (eg, systemic administration).

However, Applicants believe that if MTX is administered intradermally, MTX has similar biochemical effects (inhibits basal DNA synthesis) and biological effects (mitotic inhibition and They discovered that this can lead to the generation of damaged cells. This means that the local MT of the prior art
It is necessary that the X formulation does not allow MTX to penetrate the skin sufficiently and that the formulation contains a skin permeation reagent that facilitates the penetration of MTX into the underlying epidermal cells.

This led to subsequent discoveries. therefore.

MTX or its analogs can be administered locally in an effective manner to skin suffering from hyperproliferative epithelial diseases;
It has been discovered that the side effects associated with systemic MTX administration are thereby avoided.

Methotrexate (MTX) and its analogues effective against hyperproliferative epithelial diseases (eg, Qiantang) are referred to collectively as methotrexate or MTX. Methotrexate and its analogs are recognized by those skilled in the art as a group of agents with therapeutic activity against hyperproliferative epithelial diseases (e.g., xerotic and malignant cells) (Mc
Cullough et al (1976) J In
best Derm 66:103-107)
.

Analogs of MTX include the following compounds.

Non-limiting examples include: 3",5"-dichloromethotrexate, monoalkyl and dialkyl esters of methotrexate (eg, dimethyl or diethyl esters of methotrexate).

Mono- and dialkyl esters of 3'', 5''-dichloromethotrexate, e.g.

3',5'-dichloromethotrexate ethyl ester or 3',5'-dichloromethotrexate diethyl ester). More preferred mono- and dialkyl esters typically contain from 1 to about 10 carbon atoms per alkyl group, and often from 1 to about 5 carbon atoms per alkyl group.

The term "hyperproliferative epithelial disease" as used herein refers to a skin condition characterized by proliferation or incomplete cell differentiation of one epidermal cell. These conditions can occur spontaneously or be induced by extracorporeal means (eg, exposure to radiation or chemicals). Such diseases include xerosis, cervical hypoplasia (c
ervicaldisplasia), other premalignant trauma, fungal diseases (mycosis fungoides)
Although the treatment of dry skin is of particular relevance to the present invention, the topical compositions of the present invention are also particularly relevant.

Used to treat other hyperproliferative epithelial diseases. Such diseases include herpes simplex, viral diseases such as warts, primary skin malignancies (e.g. squamous cell carcinoma, basal cell carcinoma, black Ml) or internal malignancies on the skin. There is metastatic trauma, which exists.

A major component of the composition for topical application will be one topical carrier. The term "topical carrier" as used herein refers to a carrier material suitable for topical application of a drug such as MTX. This carrier includes any material known to cosmetologists and doctors. 1 for a suitable carrier
For example, water, liquid alcohol, liquid glycol, liquid polyalkylene glycol, liquid ester, liquid amide,
Included are liquid protein hydrolysates, liquid alkylated protein hydrolysates, liquid lanolin and lanolin derivatives, and similar materials commonly used in cosmetic and pharmaceutical compositions. Examples of carriers herein include alcohols (including both -hydric and polyhydric alcohols, such as ethanol, isopropanol, glycerol, sorbitol, 2-methoxyethanol, diethylene glycol, ethylene glycol, hexylene glycol, mannitol). and propylene glycol); ethers (e.g. diethyl ether or dipropyl ether);
propylene glycol and methoxypolyoxyethylene; carbon waxes with molecular weights ranging from 200 to 20,000; polyoxyethylene glycerol; polyoxyethylene; sorbitol; and stearoyl diacetin. Also oil-in-water emulsions like cold cream bases. Available for use. The topical carriers described herein also include various reagents and ingredients commonly used in dermatological and cosmetic ointments and lotions, such as perfumes, thickeners such as carboxymethylcellulose, stabilizers, Surfactants, emollients, colorants, and the like may be present in the carrier.

Here are topical carriers like ointments, creams, salves, jellies, pastes and lotions.

It is collectively described as a "viscous, localized drug carrier." Generally, viscous and topical drug carriers.

It is a lubricated type or a wet type. In the wet type, the carrier is mainly composed of water or alcohol, so it does not dry out the patient's skin. Examples of viscous and localized drug carriers include Eu
There is "cerin" cream.This cream is
1ersdorf Inc., Norwalk, C.
It is commercially available in N. It is a fragrance-free moisturizing formula for dry skin and contains water, petroleum, mineral oil, mineral wax, wool fat alcohol, and 2-bromo-2-nitropropane-1,3-diol. . Other viscous, localized drug carriers are known as “vehicles”.
NJ (Neutrogena Corp, Lo
sAngeles, CA)lAquaphores
phor) Ointment base (Dur Institute, 5outh
Norwalk+ CN), Unibase (Uniba
se) Ointment base (Parke-Davis, Detr.
oit+MI), and petrolatum (Petrola
tum). Other types of topical carriers are known and 1
For example, a mixture of one or more of ethanol, isopropanol, N-methylpyrrolidone (NMP) and water.

Another necessary ingredient in the topical therapeutic compositions of the present invention is a skin-penetrating reagent.

A more preferred permeation reagent is U.S. Patent No. 398,981.
1- disclosed in 6°4405616 and 4316893
Substituted-azacyclohebutan-2-one. These disclosures are set forth herein in their entirety. These compounds have the following general formula: where Rl is selected from the group consisting of H,, J-; and alkyl having 1 to 4 carbon atoms.

R is selected from the group consisting of alkyl of 1 to 18 carbon atoms, and phenyl, and n is O or 1 to 1
It is a positive integer of 0.

Examples of these include 1-methyl-azacyclohebutane-2
-one, 1-octyl-azacyclohebutan-2-one, and 1-nonyl-azacyclohebutan-2-one. A particularly preferred member of this class is 1-dodecyl-azacycloheptan-2-one, which is sold under the Azone trademark as Nelson Re5.
arch & Development Co.

Commercially available from Irvine, CA. The concentration of these permeation reagents is usually between about 0.1% and about 10%, preferably about 0.1% by weight of the local carrier.
% and about 5%.

The final dose delivered to the afflicted tissue by topical application is 9 concentrations of MTX in the topical carrier.

It depends not only on the amount of the topical composition (which is applied to the afflicted tissue), the frequency of application (also on other factors). It may range from 1% to about 10%.

Concentrations outside this range may be effective depending on other factors. However, nine topical formulations will have an MTX concentration of about 0.5%, 1%, 1.5% or 2%, based on the total weight of the composition.

Concentrations and specific topical formulations are as described above.

One is selected so that the basal epidermal cells are supplied with a therapeutically effective amount of MTX. Determination of the appropriate concentration for this skin permeation reagent as well as MTX is within the skill of one of ordinary skill in the art in view of the present disclosure.

A variety of formulations can be readily selected in vitro and in vivo as described in this example.

Particularly preferred compositions for topical application are:

A composition comprised of methotrexate and azone in a viscous, topical drug carrier. Generally, according to Azone's excellent skin penetration properties.

This formulation also allows the use of low concentrations of MTX.

More preferred formulations of the invention include about 0.1%, 0.5% or 1%
of MTX, and about 3% Azone. This gel carrier is polyethylene glycol (PEG200) which solubilizes MTX.

Isopropanol as a co-solvent for MTX.

Carbopol 934P gelling agent,
Tween 20 nonionic emulsifier,
It may contain triethanolamine to neutralize the gelling agent, and hydrochloric acid to cause MTX to exist as a combination of base and hydrochloride.

Local administration of MTX requires withdrawal or repeated applications over several days.

The exact number of applications, as well as the duration of treatment, generally.

As well as the concentration of MTX in this topical formulation.

It will also depend on the efficacy of this skin permeation reagent. for example,
A topical formulation containing about 1% methotrexate and about 3% azone should produce a therapeutic effect on a regimen of twice-daily application to distressed tissues for 3-6 weeks. It is believed that there is.

The following examples are provided for illustrative purposes only and are not intended to limit the scope of the invention in any way. Percentages are by weight unless otherwise noted.

(Example) The present invention will be described below with reference to examples.

Methotrexate was obtained from Lederle Laboratories (Pearl River, NY). Sodium methotrexate (3', 5', and 7 positions are labeled with 3H. 250 mCl/mmol) is
From Amersham, before use, 0.006M
Purified by chromatography on thin cellulose plates in potassium phosphate buffer, pH 6.0. Vehicle N is Neutrogena Corporation.

Obtained from Los Angeles, CA.

Transdermal permeation of MTX was measured in a glass diffusion cell as previously described (McCullough et al.
1976) J Invest Derm 66:
103-107) *Human skin obtained by dissecting the abdominoplasty was used in these studies. First, 10μC
0.5 d of the material containing methotrexate-3H9 and various excipients were applied to one epidermal surface, cooled to give a final concentration of 2% (0.2
rn1/cTA). A total of lOug of methotrexate was delivered to each diffusion cell. The skin reservoir contained phosphate buffered sarin. Three diffusion cells were operated for each excipient. The diffusion cell was incubated at 28°C with continuous agitation. Methotrexate permeation was followed by liquid scintillation counting at various time points up to 48 hours. After an incubation period of 48 hours, the test solution was removed and the epidermal surface was washed three times with vehicle (in which the drug being tested was dissolved). This step effectively removed all the drugs that had not penetrated the stratum corneum layer. This epidermis is 6
The skin was separated by heating for 1 minute on a heat plate at 0°C. MTX is Werkheise
r et al. (1962) J Pharmacol Exp T
using the method described in her 137:162-166.

Extracted from the epidermis and dermis. This method specifically dissociates MTX from cell-bound proteins. This layered stratum corneum was removed by repeatedly peeling with cellophane tape until the skin glowed.

The results are shown in Table 1. water (p<0.01) and n-
There was a significant increase in the permeation of MTX at both 24 and 48 hours in Vehicle N when compared to decyl methyl sulfoxide (p<0.01). Vehicle N too. compared to other excipients.

A slight increase occurred in epidermal MTX.

Table 1 Excipient permeation 124hr
48hr Month %゛ %0 μg Water 6±2 0.06 15
±5 0.1501゜MSO (2.5%) 12±10
0.12 36±28 0.36 Vehicle N
50±11 0.50 124±10 1.2
Mean ± SD for four 4 m diffusion vessels Sample C obtained after 48 h incubation % of applied dose Drug content 1·ゝ stratum corneum Epidermis μg % μg %0 9 ± 5 0.09 0.1 ± OQ ,QOL21±5
0.21 0.8±0.3 0.00816±8
0.16 1.5±0.5 0.01 s50'l Deoxyuridine (labeled with 3H at position 6) 2
5 Ci/mmol) is New England Nu
Obtained from clear. Other materials were obtained as described in Example 1.

HR5/J albino hairless mouse (2-3 months old)
was used in these experiments. To induce a deficiency of major fatty acids, 3-week-old animals were fed a diet lacking essential fatty acids (Teklad) for 60 days.

Two topical MTX preparations at various concentrations or excipient controls daily for 1 or 3 days.

O on a 1.5 cm x 3 cva area of the skin on the back.
It was applied in an amount of 0.05d. For each treatment group, 6 to
Ten mice were used. To analyze methotrexate effects on novel pathways on epidermal DNA synthesis.

Deoxyuridine containing tritium was used as a precursor. Incorporation of deoxyuridine, including tritium, into DNA is dependent on the normal action of dihydrofolate reductase, which is blocked by methotrexate. - at selected time points after multiple or repeated applications of MTX.

To measure epidermal DNA synthesis in treated back skin, tritium-containing deoxyuridine (50-
IP was injected into mice with 100 μCi),
The local effects of MTX were determined. DNA synthesis was measured simultaneously in untreated abdominal skin of animals treated with vehicle N and MTX to determine the systemic effects of MTX at separate sites. One hour after isotope administration, animals were sacrificed by dislocating their necks.

Treated back skin and untreated abdominal skin were obtained. Skin specimens were incubated in 2M sodium bromide at 37°C.
Two epidermis were removed after incubation for an hour. Halp
rin et al. (1979) J Invest De
rm 73:359-363 using the method described in
Extract the DNA.

And analyzed. The results of epidermal DNA synthesis are converted into DNA
It was calculated as a count of 7 minutes/μg. The effect of TX treatment on dorsal and abdominal DNA synthesis was expressed as % of DNA synthesis in vehicle N-treated dorsal skin and untreated abdomen in control animals.

Peakle N3 in normal hairless mouse control animals
Two consecutive applications resulted in epidermal thickening characterized by epidermal excoriation, acanthosis, and granular layer thickening. 2 of vehicle N in normal mice as well as hyperproliferative hairless mice lacking essential fatty acids.
%MTX three times daily resulted in significant epithelial atrophy in the applied areas, whereas no similar occurrence occurred in untreated abdominal skin.

Three daily applications of 2% MTX and Vehicle N reduced epithelial DNA synthesis by 50% at 54 and 72 hours in treated blue skin compared to Vehicle N treated controls, respectively. and inhibited in 75%. In this experiment, significant (p<O,oo
i) No systemic methotrexate effects were observed. However, up to 72 hours (24 hours after the last treatment) 2 applications of 0MTX 4 times daily resulted in both local and systemic effects on both the skin of the back, and the untreated skin of the abdomen. In , a global inhibition of DNA synthesis was obtained.

Even when applying 1% MTX of Vehicle N three times daily.

Inhibition of DNA synthesis occurred in the treated skin 6 hours after the last treatment. However, at low concentrations, no local or systemic effects occurred. Local MTX (0,5%) in N-decylmethyl sulfoxide is similarly
DNA synthesis does not occur in this manner.

Epidermal DNA synthesis in normal mice and in hairless mice lacking essential fatty acids is a useful bioassay for the drug activity of topical chemotherapy against dry skin (L
owe et at (1977) Br J Derm
ato196:155-162; Lowe et a
l (1981) Arch Dermatol Dan 7
:394-398. ), local MTX/Dl'I
SO preparation, which has already been shown to have no clinical activity in Qianfu. Weinstein et
al (1981) ArchDermatol
Dan 7:388-393) was found to have no effect on epidermal DNA synthesis in a hairless mouse model. In this model, MTX in Vehicle N inhibited DNA synthesis and treated the skin 6 hours after the third application, with no significant effect on the distant skin.

− Application of the drug twice is ineffective. 24 hours after the third application, DNA synthesis was inhibited in both treated and remote skin sites. This is probably due to MTX entering the mouse's systemic circulatory system.

In vitro transdermal permeation studies were conducted according to the method of Example 1 using topical gel formulations containing approximately 3% Azone and various concentrations of MTX.

The results (shown in Table 2) demonstrate a dose-dependent increase in elbow x penetration in vitro for this vehicle. It also shows that Azone is an effective permeation reagent.

(Leaving space below) The formulation of Example 3 of this study shows that D.
The effect on NA synthesis was tested. A piglet's skin is quite similar to human skin. The effect of drugs on the proliferation of epidermal cells is a recognized test for the effectiveness of therapeutic protocols against hyperproliferative skin diseases.

Piglets were collected at Charles River Research Institute, In
c, (Wilmington.

Obtained from MA). The drug preparation was applied to a small area (approximately 1 square inch) of the skin on the back according to one selected application schedule. Six consecutive hours after the last application, tritylated thymidine was injected into the drug treatment site for autoradiographic evaluation of epidermal proliferation as measured by two epidermal labeled indicators (Weinstein &
Van 5cott, (1965) J.

Invest, Derm, 44:413).

Vehicles containing various concentrations of MTX were applied daily for 7 days. tritylated deoxyuridine.

On day 7, the treated areas as well as untreated control areas were injected. DNA synthesis was measured as described in Example 2.

The results of this test are shown in Table 3. Local MTX application (
by both 0.5% and 1.0%).

Epidermal DNA synthesis was inhibited. A slight decrease in labeling intensity was also observed, reflecting a decrease in the rate of DNA synthesis compared to the excipient control.

Table 3 Estimated value of methotrexate% (%0 (RX L) 8.
1 + 1.00 (excipient) 11.2±1
．． 60.5% 0
．． Of 0.01.0%
0.8 ± 1.4] [Example 1 of MTX in vitro in various cases
Tritylated MTX was prepared using the method described in
% by weight of seed excipients were tested in vitro on one person's skin. The results are shown in Table 4 below. As can be seen, water was not an effective excipient. Vehicle N, which is an excipient containing N-methylpyrrolidone (NMP), and Azone (1-dodecyl-
Transdermal penetration was maximal at 24 hours using azacycloheptan-2-one). Epidermal content at 48 hours was also highest for these excipients.

(Left below) Transdermal penetration (μg) Wipe M1 hr 24hr 48hruz
o 1±16±2 15±5 (pH8,5
) C, oMSo 1±112±1036±28
(25%H20) Vehicle N 1±050±11 124±1ONM
P (38%) 1±127±5 46±21(E
tOH: 43%: H, 019%) Azone
1±069±23 158±60 Table 4 Laminar stratum corneum content (48 hours) Epidermal content (48 hours) ±s, o, ±S, D.

9±50 21 Sat 5 1±016±8
2±134±14
2±185±19 3±1
Further studies in the piglet model were conducted to test various carriers with different concentrations of MTX and Azone as described in Example 4. The formulation was applied daily for 7 days.

'H-UdR at 3 sites per treatment area.
was injected. Recordings were made by audiograph at the fourth week. Transdermal permeation and leaf synthesis were measured using the methods described in Examples 1 and 2.

The results are shown in Table 5. As you can see here.

A standard containing MTX at a concentration ranging from 0.5% to 1% and Azone at a concentration ranging from 3:1 to 10%.

The formulation effectively penetrates the epidermis and facilitates DNA synthesis.
Aim well. The increase in DNA synthesis observed in formulations with 1% MTX and 10% Azone was significantly increased.

Well, the irritation to the epidermis caused by the excipients.

Ru.

JLJE MTX% Isoproper Tool: H2O1% Isoproper Tool: 1% H! 0: Hydrochloric acid gel 0% gel 1% Hydrochloric acid gel 1% Hydrochloric acid gel 0.5% gel 0% Gel 1% Hydrochloric acid gel 1% Hydrochloric acid gel 0.5% Control (untreated) - Table 5 0% 10% 18. 8±8.910
% 6.8±11.8 74.310%
5.8±4.3 1067%
13.9±8.2 --7% 1.8±
3.124.5 7% 0.0±0.0 25.37%
0.8±1.48.1 3% 11.2±1.60 3% 0.5±0.9 26.13%
0.8±1, 4 25.63%
0.0±0.0 11.58.1±1.0 MTX permeation into the epidermis of various excipient formulations was evaluated in vitro at various MTX concentrations. Evaluations were made as described in Example 1. If it is not written in the number,
All excipients were gel formulations containing 35 azone. These results are shown in Table 6, and this data demonstrates that in the excipients tested, there is a dose-dependent effect on MTX permeation. Using standard formulation J containing 10% Azone, 45S Tubulobanol, 45% water:
Maximum transmission was obtained. With excipients containing 1% old, significant permeation was obtained with all excipients. These excipients all have similar permeability.

(Remains below) Table 6 Azone Gel #1 0.1 1.
4±0.3 Azone Gel #1 0.5
11.7±3.1 Azone Gel #1
1.0 24.7±3.5 Azone Gel #2
0.1 1.4±0.3 Azone Gel #2 0.5 7.9±1.5 Azone Gel #2 1.0 19.4±7
．． 9 Standard Azone: IPA: Water 1.0
PA of 84.8 ± 9.6 (10: 45: 45) 14.5 ± 3.0 58.9 ± 20.0 49.4 ± 21.7 1.3 ± 0.6 5.1 ± 2. 8 12.0 ± 4.6 71.8 ±52.0 Since it will be obvious to those skilled in the art that modifications may be made.

It is intended that the invention be limited only by the scope of the claims appended hereto.

that's all

Claims (1)

[Scope of Claims] 1. A composition for topical application to treat hyperproliferative epithelial diseases, the composition containing the following substances: (a) methotrexate to the basal epidermal cells of a mammal; a topical carrier comprising an effective amount of a skin-penetrating reagent that penetrates the skin in an amount sufficient to inhibit DNA synthesis in the underlying epidermal cells, wherein the skin-penetrating reagent has the formula: ▲ Numerical formulas, chemical formulas, tables, etc. ▼ where R' is selected from the group consisting of H and alkyl having 1 to 4 carbon atoms; ~18 alkyl, and phenyl, and n is 0 or a positive integer from 1 to 10. (b) A therapeutically effective amount of an agent selected from the group consisting of methotrexate and methotrexate analogs. 2. The drug is present in an amount of about 0.1 to
A composition according to claim 1, wherein the composition is present in an amount of about 10%. 3. The composition according to claim 1, wherein the drug is methotrexate. 4. The composition according to claim 1, wherein the drug is 3',5'-dichloromethotrexate. 5. The composition according to claim 1, wherein the drug is a monoalkyl ester or dialkyl ester of methotrexate. 6. The composition according to claim 1, wherein the skin permeation reagent is 1-dodecyl-azacycloheptane. 7. The composition according to claim 3, wherein the skin permeation reagent is 1-dodecyl-azacycloheptane. 8. The concentration of the 1-dodecyl-azacycloheptane is
8. The composition of claim 7, wherein the amount ranges from about 0.1 to about 10% by weight of said topical carrier. 9. The concentration of the 1-dodecyl-azacycloheptane is
8. The composition of claim 7, wherein the amount ranges from about 0.1 to about 5% by weight of said topical carrier. 10. Methotrexate per weight of the composition;
8. The composition of claim 7, wherein the composition is present in a concentration of about 0.1 to about 10%. 11. Methotrexate per weight of the composition;
9. The composition of claim 8, wherein the composition is present in a concentration of about 0.1 to about 10%. 12. Methotrexate per weight of the composition;
Claim 8 present in a concentration of about 0.5 to about 5%
The composition described in Section. 13. The drug is about 0.5 per weight of the composition.
10. The composition of claim 9, wherein the composition is methotrexate present at a concentration of ~5%. 14. The composition of claim 1, wherein the localized carrier is a viscous localized drug carrier. 15. The composition of claim 6, wherein the topical carrier is a viscous topical drug carrier. 16. The composition of claim 7, wherein the topical carrier is a viscous topical drug carrier. 17. The composition of claim 11, wherein the topical carrier is a viscous topical drug carrier. 18. A method for therapeutically treating a hyperproliferative epithelial disease in a mammal, comprising: a composition for topical application to treat a hyperproliferative epithelial disease; and applying the composition to a mammal suffering from the hyperproliferative epithelial disease; , a topical carrier comprising in an amount sufficient to inhibit DNA synthesis in the underlying epidermal cells, wherein the skin permeation reagent is a compound having the formula: ▲ mathematical formula, chemical formula , tables, etc. ▼ where R' is selected from the group consisting of H and alkyl having 1 to 4 carbon atoms, R is selected from the group consisting of alkyl having 1 to 18 carbon atoms, and phenyl and n is 0 or a positive integer from 1 to 10. (b) A therapeutically effective amount of an agent selected from the group consisting of methotrexate and methotrexate analogs. 19. The method of claim 18, wherein the drug is methotrexate and the skin permeation reagent is 1-dodecyl-azacycloheptane. 20. A method in which the drug is methotrexate and the skin permeation reagent is 1-dodecyl-azacycloheptane, wherein the methotrexate is about 0% by weight of the composition.
．． present in a concentration of 1 to about 10%, and the 1-dodecyl-
19. The method of claim 18, wherein the concentration of azacycloheptane ranges from about 0.1 to about 10% by weight of the topical carrier.